production and characterization of recombinant light chain and carboxyterminal heavy chain fragments of tetanus toxin

نویسندگان
چکیده

background: light chain (lc) and heavy chain carboxyterminal subdomain (hcc) fragments are the most important parts of tetanus neurotoxin (tent) which play key roles in toxicity and binding of tent, respectively. in the present study, these two fragments were cloned and expressed in a prokaryotic system and their identity was confirmed using anti-tent specific polyclonal and monoclonal antibodies. methods: lc and hcc gene segments were amplified from clostridium tetani genomic dna by pcr, cloned into pet28b(+) cloning vector and transformed in escherichia coli (e. coli) bl21(de3) expression host. recombinant proteins were then purified through his-tag using nickel-based chromatography and characterized by sds-page, western blotting and elisa techniques. results: recombinant light chain and hcc fragments were successfully cloned and expressed in (e. coli) bl21 (de3). optimization of the induction protocol resulted in production of high levels of hcc (~35% of total bacterial protein) and to lesser extends of lc (~5%). reactivity of the his-tag purified proteins with specific polyclonal and monoclonal antibodies confirmed their renatured structure and identity. conclusion: our results indicate successful cloning and production of recombinant lc and hcc fragments of tent. these two recombinant proteins are potentially useful tools for screening and monitoring of anti-tent antibody response and vaccine production.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Production and Characterization of Recombinant Light Chain and Carboxyterminal Heavy Chain Fragments of Tetanus Toxin

BACKGROUND Light chain (LC) and heavy chain carboxyterminal subdomain (HCC) fragments are the most important parts of tetanus neurotoxin (TeNT) which play key roles in toxicity and binding of TeNT, respectively. In the present study, these two fragments were cloned and expressed in a prokaryotic system and their identity was confirmed using anti-TeNT specific polyclonal and monoclonal antibodie...

متن کامل

The tetanus toxin light chain inhibits exocytosis.

The intracellular action on exocytosis of various forms of tetanus toxin was studied using adrenal medullary chromaffin cells, the membrane barrier of which has been removed by permeabilization with streptolysin O. Such cells still release catecholamines on stimulation with calcium. The two-chain form of tetanus toxin (67 nmol/l) strongly inhibited exocytosis, but only if dithiothreitol was pre...

متن کامل

synthesis and characterization of some macrocyclic schiff bases

ماکروسیکلهای شیف باز از اهمیت زیادی در شیمی آلی و دارویی برخوردار می باشند. این ماکروسیکلها با دارابودن گروه های مناسب در مکانهای مناسب می توانند فلزاتی مثل مس، نیکل و ... را در حفره های خود به دام انداخته، کمپلکسهای پایدار تولید نمایند. در این پایان نامه ابتدا یک دی آلدئید آروماتیک از گلیسیرین تهیه می شود و در مرحله بعدی واکنش با دی آمینهای آروماتیک و یا آلیفاتیک در رقتهای بسیار زیاد منجر به ت...

15 صفحه اول

Isolation, purification, and characterization of fragment B, the NH2-terminal half of the heavy chain of tetanus toxin.

Fragment B, the N-terminal half of the heavy chain, an important domain of the tetanus neurotoxin molecule, was isolated for the first time. Tetanus toxin (composed of three domains, A, B, and C) was prepared from culture filtrates. Fragment A-B, derived from the toxin treated mildly with papain, was used for the isolation of fragment B. Fragment A-B obtained was dissociated into fragments A an...

متن کامل

extraction and characterization of allium irancum plant extract and its application in the green synthesis of silver nano particles and oxidation of thiocarbony1 compounds

سنتز سبز نانوذرات فلزی (nps) درسالهای اخیر توجه بسیارزیادی را به خود جلب کرده است. زیرا این پروتوکل کم هزینه وسازگار با محیط زیست از روش های استاندارد سنتز. در این پایان نامه ما گزارش میکنیم یک روش ساده و سازگار با محیط زیست برای سنتز نانوذرات نقره با استفاده از محلول آبی عصاره گیاه allium iranicum به عنوان یک عامل کاهش دهنده ی طبیعی. نانو ذرات نقره مشخص شد با استفاده از تکنیک های uv-visible، x...

study the effects of low level laser diode on no production and healing of diabetic and cortisone acetate and normal wounded sd rats

nitric oxide (no) is a small molecule synthesized by most of mammalian cells with diverse biological activities including vasodilatation, host defense and wound healing. impaired wound healing is a common occurrence among diabetics and patients receiving glucocorticoid therapy. on the other hand the application of laser in biomedical area have been increased. thus, the current studies were desi...

15 صفحه اول

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
avicenna journal of medical biotechnology

جلد ۵، شماره ۴، صفحات ۲۲۰-۲۲۶

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023